Germline mutations of the gene encoding bone morphogenetic protein receptor 1A in juvenile polyposis.

Juvenile polyposis (JP; OMIM 174900) is an autosomal dominant gastrointestinal hamartomatous polyposis syndrome in which patients are at risk for developing gastrointestinal cancers. Previous studies have demonstrated a locus for JP mapping to 18q21.1 (ref. 3) and germline mutations in the homolog of the gene for mothers against decapentaplegic, Drosophila, (MADH4, also known as SMAD4) in several JP families. However, mutations in MADH4 are only present in a subset of JP cases, and although mutations in the gene for phosphatase and tensin homolog (PTEN) have been described in a few families, undefined genetic heterogeneity remains. Using a genome-wide screen in four JP kindreds without germline mutations in MADH4 or PTEN, we identified linkage with markers from chromosome 10q22-23 (maximum lod score of 4.74, straight theta=0.00). We found no recombinants using markers developed from the vicinity of the gene for bone morphogenetic protein receptor 1A (BMPR1A), a serine-threonine kinase type I receptor involved in bone morphogenetic protein (BMP) signaling. Genomic sequencing of BMPR1A in each of these JP kindreds disclosed germline nonsense mutations in all affected kindred members but not in normal control individuals. These findings indicate involvement of an additional gene in the transforming growth factor-beta (TGF-beta) superfamily in the genesis of JP, and document an unanticipated function for BMP in colonic epithelial growth control.

The prevalence of MADH4 and BMPR1A mutations in juvenile polyposis and absence of BMPR2, BMPR1B, and ACVR1 mutations.

BACKGROUND: Juvenile polyposis (JP) is an autosomal dominant syndrome predisposing to colorectal and gastric cancer. We have identified mutations in two genes causing JP, MADH4 and bone morphogenetic protein receptor 1A (BMPR1A): both are involved in bone morphogenetic protein (BMP) mediated signalling and are members of the TGF-beta superfamily. This study determined the prevalence of mutations in MADH4 and BMPR1A, as well as three other BMP/activin pathway candidate genes in a large number of JP patients. METHODS: DNA was extracted from the blood of JP patients and used for PCR amplification of each exon of these five genes, using primers flanking each intron-exon boundary. Mutations were determined by comparison to wild type sequences using sequence analysis software. A total of 77 JP cases were sequenced for mutations in the MADH4, BMPR1A, BMPR1B, BMPR2, and/or ACVR1 (activin A receptor) genes. The latter three genes were analysed when MADH4 and BMPR1A sequencing found no mutations. RESULTS: Germline MADH4 mutations were found in 14 cases (18.2%) and BMPR1A mutations in 16 cases (20.8%). No mutations were found in BMPR1B, BMPR2, or ACVR1 in 32 MADH4 and BMPR1A mutation negative cases. DISCUSSION: In the largest series of JP patients reported to date, the prevalence of germline MADH4 and BMPR1A mutations is approximately 20% for each gene. Since mutations were not found in more than half the JP patients, either additional genes predisposing to JP remain to be discovered, or alternate means of inactivation of the two known genes are responsible for these JP cases.

Thymoquinone suppresses expression of inducible nitric oxide synthase in rat macrophages.

The objective of the present study was to determine the immunomodulatory role of thymoquinone (TQ) regarding its effect on the production of nitric oxide (NO) by rat peritoneal macrophages. Under certain conditions, macrophagesand certain other cells can produce high concentrations of NO from its precursor L-arginine via inducible nitricoxide synthase (iNOS)pathway. TQ has been established as the major component of the oil extracted from Nigella saliva plant seeds, which is being used frequently in herbal medicine. TQ (IC50 1.4-2.76 microM) dose- and time-dependently reduced nitrite production, a parameter for NO synthesis, in supematants of lipopolysaccharide (LPS)-stimulated (5 microg/ml) macrophages without affecting the cell viability. The protein level of iNOS in peritoneal macrophages was also decreased by TQ in a concentration-dependent manner. In addition, TQ inhibited the increase in iNOS mRNA expression induced by LPS indicated by reverse transcription-polymerase chain reaction (RT-PCR). These inhibitory effects of TQ were confirmed by immunofluorescence staining of iNOS in macrophages which showed decreased immunoreactivity for iNOS after treatment with TQ if compared with the control LPS-stimulated cells. These results suggest that TQ suppresses the production of NO by macrophages; an effect which may be useful in ameliorating the inflammatory and autoimmune conditions.

Disposition kinetics of gentamicin in normal and endometritic cows using a microbiological assay.

Gentamicin concentrations in serum, urine and milk were assayed microbiologically after intramuscular and intrauterine administrations in normal and endometritic cows. Following intramuscular injections of 5 mg gentamicin/kg b. wt. 3 times daily for three consecutive days, the highest serum concentrations occurred 1 h post administration of each dose with absorption half-lives [t0.5(ab)] ranging from 0.23 to 0.30 h for normal cows and from 0.21 to 0.29 h for endometritic cows. The elimination half-lives [t0.5(beta)] ranged from 2.51 to 2.95 h (normal cows) and from 2.71 to 3.29 h (endometritic cows). Following intrauterine administration of 4 mg gentamicin/kg. b. wt. once daily for three consecutive days, the drug peaked in serum 2 h after each dose with [t0.5(ab)] ranging from 0.47 to 0.52 h (normal cows) and from 0.57 to 0.68 h (diseased cows), while the drug was eliminated faster in endometritic cows than in normal cows. The mean systemic bioavailability were (70%) and (30%) after intramuscular and intrauterine administration, respectively. To compare serum concentrations, gentamicin was assayed in urine and milk in high and low concentrations, respectively.

Disposition kinetics of cephradine in normal and Escherichia coli infected goats.

The pharmacokinetics of cephradine was studied following single and repeated intramuscular injections in normal and Escherichia coli infected goats. Bioavailability of cephradine was determined in normal goats after a single intramuscular dose. The serum concentrations of cephradine following a single and repeated intramuscular administration of 10 mg/kg b.wt. twice daily for five consecutive days, peaked 2 hours after each intramuscular dose with a lower significant value recorded in E. coli infected goats than in normal goats. The absorption half-lives (t0.5(ab)) following a single intramuscular injection of cephradine was significantly higher in E. coli infected goats (1.18 h) than in normal goats (0.64 h). The elimination half-lives (t0.5(beta)) of cephradine were significantly higher in E. coli infected goats than in normal goats following the administration of fifth and ninth doses. The urine and milk concentrations of cephradine were significantly lower in E. coli infected goats than in normal goats. The mean systemic bioavailability of cephradine following a single intramuscular injection in normal goats was 73.9%.

Blood and tissue concentrations of trimethoprim following its administration alone and in combination with josamycin.

Following a single oral dose of trimethoprim (10 mg/kg b. wt.) in normal fowls, the highest serum concentration achieved 4 hours post-administration with value of 0.64 microgram/ml. The absorption half-life time was 0.64 hours. The elimination half life was 4.73 hours. During repeated oral administration of 10 mg/kg b. wt., once daily for five consecutive days, trimethoprim peaked in serum, 4 h after each dose. Trimethoprim persisted in all fowl's tissues for 96 hours after stopping of drug administration. After oral administration of josamycin (18 mg/kg b. wt.) and trimethoprim (10 mg/kg b. wt.) in normal fowls, a maximum serum concentration of trimethoprim was recorded at 2 hours with half-life of absorption (t0.5(ab)) valued 0.74 hour. The elimination half-life (t0.5 beta) was 4.37 hours. During repeated oral administration of josamycin (18 mg/kg b. wt.) and trimethoprim (10 mg/kg b. wt.) once daily for five consecutive days in normal fowls, the highest plasma concentrations of trimethoprim occurred 2 hours post each dose. The daily maximum plasma concentrations during the repeated oral administration of both tested drugs were nearly constant.

Pharmacokinetics and tissue residues of josamycin in fowls.

Josamycin is a macrolide antibiotic which is produced by fermentation of cultures of Streptomyces narbonensis. It was once administrated (18 mg/kg b. wt.) in fowls via intravenous, oral and intramuscular routes for determination of blood concentration, kinetic behaviour and bioavailability. Following a single intravenous injection, the blood concentration-time-curve indicated a two compartments open model with an elimination half life value (t1/2 beta) of 1.83 +/- 0.06 hours. Both oral and intramuscular routes showed higher values, i.e. 2.33 +/- 0.13 and 2.85 +/- 0.17 hours. The lower apparent volume of distribution of Josamycin in fowls than one liter/kg elucidate higher distribution in blood than in tissues. Systemic bioavailability after both oral and intramuscular administration, i.e. 33.88 +/- 2.4 and 27.28 +/- 1.46% respectively, showed lower absorption from site of i.m. application. Josamycin was administered (18 mg/kg b. wt.) intramuscularly and orally once daily for 5 consecutive days. The drug peaked in serum 1 hour (intramuscular) and 2 hours (orally) after each dose. The recorded results revealed that serum level of Josamycin was higher after oral application (29.98 +/- 1.92 micrograms/ml) than after i.m. application. The drug persisted in the lung tissues and fat for 72 hours after administration and disappeared from all body tissues 96 hours after the last dose of repeated administration.

Serum concentrations and tissue residues of spectinomycin in chickens.

Following a single intramuscular injection of 40 mg spectinomycin/kg.b.wt. in normal chickens, a maximum serum concentration was recorded at one hour, with half-lives of absorption [t0.5(ab)] and elimination [t0.5(beta)] valued with 0.21 h and 3.27 h respectively. Following a single intravenous injection of 40 mg spectinomycin/kg b. wt. in normal chickens, the drug obeyed a three compartments open model. The mean systemic bioavailability following intramuscular injection was 3.72 %. The highest serum concentration of spectinomycin was achieved after one hour post each intramuscular dose during multiple dosage regimen. Serum and tissue concentrations of spectinomycin in slaughtered normal chickens following repeated intramuscular administration, three times daily for five consecutive days were investigated. In the present study, spectinomycin was bound in vitro with normal chicken serum protein at a level equal to 5.4 %.

Kinetic behaviour of sulphaquinoxaline and amprolium in chickens.

The pharmacokinetics of sulphaquinoxaline and amprolium hydrochloride were studied in Hubbard broiler chickens. Single doses of sulphaquinoxaline (100 mg/kg b. wt.), and amprolium hydrochloride (30 mg/kg b. wt.) were administered orally and intravenously to the same birds with 15 days interval between treatments. Sulphaquinoxaline and amprolium HCl were determined colorimetrically. Following i.v. administration, the concentration-time curve of sulphaquinoxaline and amprolium could be explained by a two compartments open model with a t1/2 alpha of 0.16 +/- 0.008 h; 0.17 +/- 0.09 h; t1/2 beta of 12.6 +/- 0.32 h, 4.89 +/- 0.3 h respectively. The total body clearance were 0.278 +/- 0.013 ml/kg/min; 0.562 +/- 0.015 ml/kg/min; volume of distribution at steady state were 0.44 +/- 0.009 L/kg, 0.34 +/- 0.005 L/kg and systemic bioavailability following oral administration were 72.65 +/- 3.38, 66.09 +/- 4.9 percent for sulphaquinoxaline and amprolium HCl respectively. Following oral administration of sulphaquinoxaline and amprolium (the same previous doses) the peak plasma concentrations (Cmax) were 107.8 +/- 1.49 micrograms/ml; 42.9 +/- 1.11 micrograms/ml and occurred at 5.56 +/- 0.1 h, 3.67 +/- 0.05 h respectively. Pharmacokinetic parameters after repeated oral daily administrations of sulphaquinoxaline and amprolium revealed that the Cmax was 184 +/- 1.02 micrograms/ml, and 55.19 +/- 0.35 micrograms/ml at 7.36 +/- 0.18 h and 5.17 +/- 0.15 h and the biological half lives were 1.67 +/- 0.057 h and 1.11 +/- 0.14 h respectively. Sulphaquinoxaline and its N4 acetyl metabolite disappeared from all body tissues at 120 hours, however amprolium persisted in most tissues for 72 hours after the last dose of repeated administrations.

Pharmacokinetics and tissue residues of netilmicin and vancomycin in chickens.

Following a single intravenous injection of 10 mg netilmicin or 30 mg vancomycin/kg b.w. to normal chickens, the tested drugs obeyed a two or three compartments open model, with half-lives of distribution of 0.30 and 0.20 hours, respectively. The elimination half-lives following intravenous injection were 0.41 and 4.85 hours, respectively. Following a single intramuscular injection of 10 mg netilmicin or 30 mg vancomycin/kg b.w. to normal chickens, the serum drug concentrations peaked 2 hours post-injection with half-lives of absorption equal to 0.63 and 0.56 hours, respectively. The mean systemic bioavailability of netilmicin and vancomycin following a single intramuscular administration in normal chickens was 27.29 and 43.61%, respectively. These values indicated a limited and moderate absorption for netilmicin and vancomycin from intramuscular site, respectively. During repeated intramuscular administrations of both antibiotics for five consecutive days in normal chickens, the serum drug concentrations peaked two hours post each dose. The drugs concentrations during multiple dosage regimens were significantly increased in 2nd, 3rd, 4th and 5th days in comparison to the 1st day. Netilmicin and vancomycin persisted in liver and kidney for 96 and 120 hours, respectively, after the last intramuscular administration. The withdrawal time of netilmicin and vancomycin could be considered as five and six days, respectively. The in-vitro protein binding percents of netilmicin and vancomycin were 7.45 and 5.81%, respectively.

Some pharmacodynamic and biochemical aspects of cefamandole.

The pharmacodynamic and nephrotoxic effects of cefamandole were investigated. Cefamandole at concentrations of 512 and 1024 micrograms/ml bath caused complete relaxation in isolated guinea pig ileum and rabbit duodenum, respectively. Concentrations of 2048 and 4096 micrograms cefamandole/ml bath caused marked stimulation in force and frequency of rat uterine muscle in all stages of sex cycle. Cefamandole in all tested concentrations did not induce any response on isolated guinea pig tracheal chain or isolated rabbit aortic strip. Cefamandole in concentrations of 256 to 1024 micrograms/ml bath as well as 256 and 512 micrograms/ml cannula produced marked inhibition on isolated guinea pig auricles and rabbit heart, respectively. The effect of graded increased concentrations on isolated frog gastrocnemius muscle, frog rectus abdominis muscle and rat phrenic nerve hemidiaphragm was recorded. Cefamandole in a dose of 53.2 mg/kg b. wt. in anaesthetized dogs caused very marked hypotensive effects and decrease in rate of respiration. Single intramuscular injection of cefamandole in a therapeutic (23.3 mg/kg b. wt.) and double therapeutic (46.6 mg/kg.b. wt.) doses in rabbits had no effect on electrocardiographic parameters among a period of 8 hours after injection. Effects of cefamandole on serum and urine concentrations of creatinine, urea, sodium, potassium, calcium, glucose and protein as well as clearance tests were investigated in rats.

Effects of some beta-adrenergic blockers on male fertility parameters in rats.

The effects of atenolol (9 and 18 mg/kg.b.wt.), metoprolol (3.5 and 7 mg/kg.b.wt.) and propranolol (7.5 and 15 mg/kg.b.wt.) on male rate fertility were investigated following repeated oral administrations for 60 consecutive days. Repeated administrations of atenolol (9 and 18 mg/kg.b.wt.) induced non significant effects on weights of testes, epididymis and seminal vesicle at the first day, 30 and 60 days after the last repeated oral administration for 60 days. Administration of atenolol (9 and 18 mg/kg.b.wt.), metoprolol (3.5 and 7 mg/kg.b.wt.) and propranolol (15 mg/kg.b.wt.) to male rats induced significant decrease in percent of progressive motility of sperm at first day after the last oral administration for 60 days. Atenolol (18 mg/kg.b.wt.), metoprolol (7 mg/kg.b.wt.) and propranolol (7.5 and 15 mg/kg.b.wt.) induced significant increase in sperm head and tail abnormalities at first day after the last repeated dose. All rats treated with atenolol (9 mg/kg.b.wt.), metoprolol (7.5 and 15 mg/kg.b.wt.) and propranolol (7.5 and 15 mg/kg.b.wt.) induced significant decrease in the level of testosterone hormone at first and 30 days after the last dose. Repeated administrations of atenolol, metoprolol and propranolol in therapeutic and double therapeutic doses for 60 days induced nearly similar histopathological alterations in testis, epididymis and seminal vesicles. The induced hazard effects by the tested drugs on the male rat fertility were reversible as they returned to normal values 60 days after discontinuation of therapy.

Pharmacokinetic interpretation of some antibiotics in camels.

Evaluation of the pharmacokinetic properties of 4 antibiotics: penicillin-G (P), streptomycin (S), chloramphenicol (C) and oxytetracycline (O) was performed in groups of camels following a single i.m. injection of therapeutic doses, i.e. 6000 IU, 10, 4 and 3 mg/kg b.wt., respectively. The concentrations of these antibiotics in serum were determined by microbiological assay methods. The highest serum concentrations were reached after 0.42, 1.44, 4.02 and 0.94 hr for P, S, C, and O respectively with corresponding t 1/2 alpha values of 0.12, 0.28, 1.48 and 0.17 hr and t 1/2 beta values of 1.09, 8.28, 6.20 and 7.00 hr.